Phosphorylation of Spinophilin Modulates Its Interaction with Actin Filaments

Spinophilin is a protein phosphatase 1 (PP1)- and actin-binding protein that modulates excitatory synaptic transmission and dendritic spine morphology. We report that spinophilin is phosphorylated in vitro by protein kinase A (PKA). Phosphorylation of spinophilin was stimulated by treatment of neostriatal neurons with a dopamine D1 receptor agonist or with forskolin, consistent with spinophilin being a substrate for PKA in intact cells. Using tryptic phosphopeptide mapping, site-directed mutagenesis, and microsequencing analysis, we identified two major sites of phosphorylation, Ser-94 and Ser-177, that are located within the actin-binding domain of spinophilin. Phosphorylation of spinophilin by PKA modulated the association between spinophilin and the actin cytoskeleton. Following subcellular fractionation, unphosphorylated spinophilin was enriched in the postsynaptic density, whereas a pool of phosphorylated spinophilin was found in the cytosol. F-actin co-sedimentation and overlay analysis revealed that phosphorylation of spinophilin reduced the stoichiometry of the spinophilin-actin interaction. In contrast, the ability of spinophilin to bind to PP1 remained unchanged. Taken together, our studies suggest that phosphorylation of spinophilin by PKA modulates the anchoring of the spinophilin-PP1 complex within dendritic spines, thereby likely contributing to the efficacy and plasticity of synaptic transmission

Quantile regression of tobacco tax pass-through in the UK 2013–2019. How have manufacturers passed through tax changes for different tobacco products?

Background: The effectiveness of tax increases relies heavily on the tobacco industry passing on such increases to smokers (also referred to as ‘pass-through’). Previous research has found heterogeneous levels of tax pass-through across the market segments of tobacco products available to smokers. This study uses retail sales data to assess the extent to which recent tax changes have been passed on to smokers and whether this varies across the price distribution. Methods: We use panel data quantile regression analysis on Nielsen commercial data of tobacco price and sales in the UK from January 2013 to March 2019 combined with official UK tax rates and inflation to calculate the rate of tax pass-through for factory made (FM) cigarettes and roll your own (RYO) tobacco. Results: Following increases in the specific tax payable on tobacco, we find evidence of overshifting across the price distribution for both FM and RYO. The rate of the overshift in tax increased the more expensive the products were. This was consistent for FM and RYO. Additionally, our findings suggest that the introduction of standardised packaging was not followed by changes in how the tobacco industry responded to tax increases. Conclusions: Following the repeated introduction of increases in specific tobacco tax as well as standardised packaging, we show that the tobacco industry applies techniques to keep the cheapest tobacco cheaper relative to the more expensive products when passing on tax increases to: smokers

The JCMT Nearby Galaxies Legacy Survey -- XI. -- Environmental Variations in the Atomic and Molecular Gas Radial Profiles of Nearby Spiral Galaxies

This article has been accepted for publication in Monthly Notices of the Royal Astronomical Society. ©: 2017 The Author (s). Published by Oxford University Press on behalf of the Royal Astronomical Society. All rights reserved.We present an analysis of the radial profiles of a sample of 43 HI-flux selected spiral galaxies from the Nearby Galaxies Legacy Survey (NGLS) with resolved James Clerk Maxwell Telescope (JCMT) CO $J=3-2$ and/or Very Large Array (VLA) HI maps. Comparing the Virgo and non-Virgo populations, we confirm that the HI disks are truncated in the Virgo sample, even for these relatively HI-rich galaxies. On the other hand, the H$_{2}$ distribution is enhanced for Virgo galaxies near their centres, resulting in higher H$_{2}$ to HI ratios and steeper H$_{2}$ and total gas radial profiles. This is likely due to the effects of moderate ram pressure stripping in the cluster environment, which would preferentially remove low density gas in the outskirts while enhancing higher density gas near the centre. Combined with H$\alpha$ star formation rate data, we find that the star formation efficiency (SFR/H$_{2}$) is relatively constant with radius for both samples, but Virgo galaxies have a $\sim40\%$ lower star formation efficiency than non-Virgo galaxies.Peer reviewedFinal Published versio

The JCMT Nearby Galaxies Legacy Survey – XI. Environmental Variations in the Atomic and Molecular Gas Radial Profiles of Nearby Galaxies

We present an analysis of the radial profiles of a sample of 43 H I-flux selected spiral galaxies from the Nearby Galaxies Legacy Survey (NGLS) with resolved James Clerk Maxwell Telescope (JCMT) CO J = 3 − 2 and/or Very Large Array (VLA) H I maps. Comparing the Virgo and non-Virgo populations, we confirm that the H i discs are truncated in the Virgo sample, even for these relatively H i-rich galaxies. On the other hand, the H2 distribution is enhanced for the Virgo galaxies near their centres, resulting in higher H2 to H I ratios and steeper H2 and total gas radial profiles. This is likely due to the effects of moderate ram pressure stripping in the cluster environment, which would preferentially remove low-density gas in the outskirts while enhancing higher density gas near the centre. Combined with Hα star formation rate data, we find that the star formation efficiency (SFR/H2) is relatively constant with radius for both samples, but the Virgo galaxies have an ∼40 per cent lower star formation efficiency than the non-Virgo galaxies

A core outcome set for seamless, standardized evaluation of innovative surgical procedures and devices (COHESIVE):a patient and professional stakeholder consensus study

To develop a COS, an agreed minimum set of outcomes to measure and report in all studies evaluating the introduction and evaluation of novel surgical techniques. SUMMARY OF BACKGROUND DATA: Agreement on the key outcomes to measure and report for safe and efficient surgical innovation is lacking, hindering transparency and risking patient harm. METHODS: (I) Generation of a list of outcome domains from published innovation-specific literature, policy/regulatory body documents, and surgeon interviews; (II) Prioritization of identified outcome domains using an international, multi-stakeholder Delphi survey; (III) Consensus meeting to agree the final COS. Participants were international stakeholders, including patients/public, surgeons, device manufacturers, regulators, trialists, methodologists, and journal editors. RESULTS: A total of 7972 verbatim outcomes were identified, categorized into 32 domains, and formatted into survey items/questions. Four hundred ten international participants (220 professionals, 190 patients/public) completed at least one round 1 survey item, of which 153 (69.5%) professionals and 116 (61.1%) patients completed at least one round 2 item. Twelve outcomes were scored “consensus in” (“very important” by ≥70% of patients and professionals) and 20 “no consensus.” A consensus meeting, involving context: modifications, unexpected disadvantages, device problems, technical procedure completion success, patients’ experience relating to the procedure being innovative, surgeons’/operators’ experience. Other domains relate to intended benefits, whether the overall desired effect was achieved and expected disadvantages. CONCLUSIONS: The COS is recommended for use in all studies before definitive randomized controlled trial evaluation to promote safe, transparent, and efficient surgical innovation

Estimation of integrated price elasticities for alcohol and tobacco in the United Kingdom using the living costs and food survey 2006–2017

Introduction Evidence shows that price is an important policy lever in reducing consumption of alcohol and tobacco. However, there is little evidence of the cross-price effect between alcohol and tobacco. Methods This paper uses an econometric model which estimates participation and consumption elasticities, on data from the UK Living Costs and Food Survey 2006–2017 and extends the literature by, for the first time, estimating joint price elasticities for disaggregated alcohol and tobacco products. This paper presents new price elasticities and compares them to the existing literature. Results The own-price elasticity estimates are all negative for both participation and consumption. There is no pattern to the estimates of cross-price elasticities. The elasticity estimates, when used in the Sheffield Tobacco and Alcohol Policy Model, produce bigger changes in consumption for the same change in price compared to other elasticity estimates in the existing literature. Discussion and Conclusions Consumption of alcohol and tobacco are affected by the prices of one another. Policymakers should bear this in mind when devising alcohol or tobacco pricing policies

Do dual purchasers behave differently? An analysis of purchasing data for households that buy both alcohol and tobacco in the United Kingdom

Background and Aims Dual purchasers of alcohol and tobacco are at increased health risk from the interacting health impacts of alcohol and tobacco use. They are also at financial risk from exposure to the dual financial cost of policies that increase alcohol and tobacco prices. Understanding whose alcohol and tobacco use exposes them to these health and financial risks is important for understanding the inequality impacts of control policies. This study explores the extent to which household spending on alcohol and tobacco combined varies between socio‐economic groups and compares this with results for households which purchase only one of the commodities. Design Cross‐sectional analysis of household‐level alcohol and tobacco purchasing data. Setting United Kingdom, 2012–17. Participants/Cases A total of 26 021 households. Measurements We analysed transaction‐level data from individual 14‐day spending diaries in the Living Cost and Food Survey (LCFS). We used this to calculate expenditure, volumes of alcohol and tobacco purchased, and the price paid per unit of alcohol (1 unit = 8 g) and per stick of tobacco. This was compared with equivalized total expenditure and quintiles of equivalized household income. Prices were calibrated and pack sizes were imputed using empirical sales data from Nielsen/CGA to correct for reporting bias. Findings Dual purchasing households spent [95% confidence interval] more on alcohol and more on tobacco than their single‐purchasing counterparts. In general, lower‐income households spent less on both alcohol and tobacco than higher‐income households. Furthermore, dual purchasing households in the lowest income group were most exposed to potential increases in price than were other income groups, with (CI = 12.41–13.15%) of their total household budget spent on alcohol and tobacco. Conclusions Dual purchasers of alcohol and tobacco in the United Kingdom appear to be concentrated evenly among income groups. However, dual purchasers may experience particularly large effects from pricing policies, as they spend a substantially higher proportion of their overall household expenditure on alcohol and tobacco than do households that purchase only one of the commodities

The effects of feed restriction, time of day and time since feeding on behavioral and physiological indicators of hunger in broiler breeder hens

Broiler breeder chickens are commercially feed restricted to slow their growth and improve their health and production, however, there is research demonstrating that this leads to chronic hunger resulting in poor welfare. A challenge in these studies is to account for possible daily rhythms or the effects of time since last meal on measures relating hunger. To address this, we used 3 feed treatments: AL (ad libitum fed), Ram (restricted, fed in the morning), and Rpm (restricted, fed in the afternoon) to control for diurnal effects. We then conducted foraging motivation tests and collected home pen behavior and physiological samples at 4 times relative to feeding throughout a 24-h period. The feed treatment had the largest influence on the data, with AL birds weighing more, having lower concentrations of plasma NEFA, and mRNA expression of AGRP and NPY alongside higher expression of POMC in the basal hypothalamus than Ram or Rpm birds (P &lt; 0.001). R birds were more successful at and had a shorter latency to complete the motivation test, and did more walking and less feeding than AL birds in the home pen (P &lt; 0.01). There was little effect of time since last meal on many measures (P &gt; 0.05) but AGRP expression was highest in the basal hypothalamus shortly after a meal (P &lt; 0.05), blood plasma NEFA was higher in R birds just before feeding (P &lt; 0.001) and glucose was higher in Ram birds just after feeding (P &lt; 0.001), and the latency to complete the motivation test was shortest before the next meal (P &lt; 0.05). Time of day effects were mainly found in the difference in activity levels in the home pen when during lights on and lights off periods. In conclusion, many behavioral and physiological hunger measures were not significantly influenced by time of day or time since the last meal. For the measures that do change, future studies should be designed so that sampling is balanced in such a way as to minimize bias due to these effects.</p

Reanalysis of single-cell RNA sequencing data does not support herpes simplex virus 1 latency in non-neuronal ganglionic cells in mice

Most individuals are latently infected with herpes simplex virus type 1 (HSV-1), and it is well-established that HSV-1 establishes latency in sensory neurons of peripheral ganglia. However, it was recently proposed that latent HSV-1 is also present in immune cells recovered from the ganglia of experimentally infected mice. Here, we reanalyzed the single-cell RNA sequencing (scRNA-Seq) data that formed the basis for that conclusion. Unexpectedly, off-target priming in 3’ scRNA-Seq experiments enabled the detection of non-polyadenylated HSV-1 latency-associated transcript (LAT) intronic RNAs. However, LAT reads were near-exclusively detected in mixed populations of cells undergoing cell death. Specific loss of HSV-1 LAT and neuronal transcripts during quality control filtering indicated widespread destruction of neurons, supporting the presence of contaminating cell-free RNA in other cells following tissue processing. In conclusion, the reported detection of latent HSV-1 in non-neuronal cells is best explained using compromised scRNA-Seq datasets. IMPORTANCE Most people are infected with herpes simplex virus type 1 (HSV-1) during their life. Once infected, the virus generally remains in a latent (silent) state, hiding within the neurons of peripheral ganglia. Periodic reactivation (reawakening) of the virus may cause fresh diseases such as cold sores. A recent study using single-cell RNA sequencing (scRNA-Seq) proposed that HSV-1 can also establish latency in the immune cells of mice, challenging existing dogma. We reanalyzed the data from that study and identified several flaws in the methodologies and analyses performed that invalidate the published conclusions. Specifically, we showed that the methodologies used resulted in widespread destruction of neurons which resulted in the presence of contaminants that confound the data analysis. We thus conclude that there remains little to no evidence for HSV-1 latency in immune cells.</p